1.Shashen Maidong Tang Enhances Efficacy of Chemotherapy in Mouse Model of Lewis Lung Cancer by Modulating JAK2/STAT3 Signaling Pathway
Lin YU ; Yaoyao WANG ; Limin LIU ; Zuowei HU ; Yanping ZHOU ; Shang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):1-10
ObjectiveTo predict the mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer via network pharmacology and validate the prediction results in animal experiments. MethodsThe potential mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer was predicted by network pharmacology, liquid chromatography-mass spectrometry (LC-MS), and molecular docking methods. C57/BL6 mice were assigned into normal, model, cisplatin, and Shasheng Maidong Tang+cisplatin groups. In addition to the normal group, the remaining groups were injected subcutaneously with 0.2 mL of 1×107 cells·mL-1 Lewis lung cancer cells to establish the Lewis lung cancer model. The daily gavage dose of Shasheng Maidong Tang was 3.58 g·kg-1, and the concentration of cisplatin intraperitoneally injected on every other day was 2 mg·kg-1. Drugs were administered for 14 d. The changes in the tumor volume and the rate of tumor suppression were monitored, and the tumor histopathological changes were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay was employed to measure the interleukin (IL)-6 and interferon (IFN)-γ levels in peripheral blood. Real-time PCR was performed to quantify the mRNA levels of Janus kinase 2 (JAK2), signal transducer and activator of transcription 1 (STAT1), and signal transducer and activator of transcription 3 (STAT3) in the tumor tissue of mice. Western blot was employed to determine the protein levels of JAK2, STAT3, B-cell lymphoma-2 (Bcl-2), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and Pim-1 proto1 (PIM1) in the tumor tissue. Immunohistochemistry was employed to detect the expression of Bcl-2 and PIM1 in the tumor tissue. ResultsNetwork pharmacological predictions indicated that Shasheng Maidong Tang might enhance the efficacy of chemotherapy for lung cancer by regulating nitrogen metabolism, AGE-RAGE signaling pathway, cancer pathway, and JAK/STAT signaling pathway. The experimental results demonstrated that tumor volume in the cisplatin group and Shasheng Maidong Tang+cisplatin group was reduced compared with the model group, with statistically distinct differences observed on days 14, 17, 20 post modeling (P<0.05). Notably, the Shasheng Maidong Tang+cisplatin therapy further decreased tumor volume compared with the cisplatin group, showing marked reductions on days 17 and 20 (P<0.05), consistent with trends visualized in tumor volume comparison charts. The Shasheng Maidong Tang+cisplatin group exhibited higher tumor inhibition rate than the cisplatin group (P<0.05). Histopathological analysis via HE staining revealed that the tumors in the model group displayed frequent nuclear mitosis, densely arranged cells, hyperchromatic nuclei, and no necrosis. Cisplatin treatment induced partial necrosis and vacuolization, while the Shasheng Maidong Tang+cisplatin group exhibited extensive necrotic regions, maximal vacuolization, disarranged tumor cells, and minimal mitotic activity. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed elevated level of IFN-γ (P<0.01) and declined level of IL-6 (P<0.01) in the peripheral blood. Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented elevated level of IFN-γ (P<0.01) and lowered level of IL-6 (P<0.01) in the peripheral blood. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin groups showed down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level STAT1 (P<0.01). Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level of STAT1 (P<0.01). Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), and STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). Compared with the cisplatin group, Shasheng Maidong Tang+cisplatin group presented down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). The Bcl-2 and PIM1 expression results obtained by immunohistochemistry were consistent with those of Western blot. ConclusionShasheng Maidong Tang may enhance the efficacy of chemotherapy in the mouse model of Lewis lung cancer by regulating the JAK2/STAT3 signaling pathway.
2.Ethical issues and countermeasures of off-label drug use in children
Limin LI ; Haipeng HU ; Xiaoqin CAO ; Xiaohong LIU
Chinese Medical Ethics 2025;38(2):227-231
Children, as a special group, frequently experience of off-label drug use worldwide. Common reasons for off-label drug use in children include the lack of data on pediatric patients during the clinical trial stage of drug development, delayed updates to drug instructions, and the non-standard professional behavior of some doctors. Off-label drug use in children is a double-edged sword. It could save lives and provide a way to explore additional functions of drugs, while it may also lead to the phenomenon of hyper-indication abuse, increasing the risk of adverse drug events. Regulating off-label drug use in children can safeguard the best treatment rights and interests of children. It is recommended to encourage pharmaceutical enterprises to conduct research and development of pediatric new drugs, simplify the approval process for drug instructions amendments, accumulate evidence-based medical evidence for off-label drug use in children, standardize the process of off-label drug use in children in medical institutions, continuously improve the standardized diagnosis and treatment capabilities of pediatricians, and actively cooperate with the families of pediatric patients in diagnosis and treatment, so as to comprehensively safeguard the rights and interests of both doctors and patients.
3.Determination of 17 types of phthalates in drinking water by solid-phase extraction coupled with multiple reaction monitoring of gas chromatography-mass spectrometry
SHI Mengxing ; YANG Yan ; DU Limin
Journal of Preventive Medicine 2025;37(1):102-108
Objective:
To determine 17 types of phthalic acid esters (PAEs) in drinking water using solid-phase extraction coupled with gas chromatography-mass spectrometry (GC-MS) in multiple reaction monitoring (MRM) mode.
Methods:
One litre of commercially available bottled water was purified using an HLB solid-phase extraction column, and was eluted with ethyl acetate, dichloromethane and average methanol. Seventeen types of PAEs were detected using a triple quadrupole gas chromatography-mass spectrometer in MRM mode. By optimizing the temperature programming and adjusting the mass spectrometry collision energy, collection efficiency was improved and matrix interference was reduced. The precision and accuracy of this method were assessed by determining the standard curves, detection limits, quantification limits, relative standard deviations (RSD) and average spiked recovery rates for the 17 types of PAEs.
Results:
The 17 types of PAEs showed good linear relationships between mass concentration and chromatographic peak areas in the range of 0.02 to 1.0 mg/L, with correlation coefficients all greater than 0.999 1. The detection limits ranged from 0.002 9 to 0.009 7 mg/kg, the quantification limits ranged from 0.008 7 to 0.029 1 mg/kg, the RSD ranged from 0.8% to 3.0%, and the average spiked recovery rates ranged from 88.8% to 111.8%.
Conclusion
Solid-phase extraction coupled with MRM of GC-MS can better determine low concentrations of PAEs in drinking water.
4.Dipsacus asper Treats Alzheimer's Disease in Caenorhabditis elegans by Regulating PPARα/TFEB Pathway
Mengmeng WANG ; Jianping ZHAO ; Limin WU ; Shuang CHU ; Yanli HUANG ; Zhenghao CUI ; Yiran SUN ; Pan WANG ; Hui WANG ; Zhenqiang ZHANG ; Zhishen XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):104-114
ObjectiveTo investigate the anti-Alzheimer's disease (AD) effect of Dipsacus asper(DA) in the Caenorhabditis elegans model, and decipher the underlying mechanism via the peroxisome proliferator-activated receptor α (PPARα)/transcription factor EB (TFEB) pathway. MethodsFirst, transgenic AD C. elegans individuals were assigned into the blank control, model, positive control (WY14643, 20 µmol·L-1), and low-, medium-, and high-dose (100, 200, and 400 mg·L-1, respectively) DA groups. The amyloid β-42 (Aβ42) formation in the muscle cells, the paralysis time, and the deposition of amyloid β-protein (Aβ) in the head were detected. The lysosomal autophagy in the BV2 cell model was examined by Rluc-LC3wt/G120A. The expression levels of lysosomal autophagy-related proteins LC3Ⅱ, LC3I, LAMP2, and TFEB were detected by Western blot. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA levels of autophagy-related genes beclin1 and Atg5 and lysosome-related genes LAMP2 and CLN2 downstream of PPARα/TFEB. A reporter gene assay was used to detect the transcriptional activities of PPARα and TFEB. Immunofluorescence was used to detect the fluorescence intensity of PPARα, and the active components of the ethanol extract of DA were identified by UPLC-MS. RCSB PDB, Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and Autodock were used to analyze the binding between the active components and PPARα-ligand-binding domain (LBD). ResultsCompared with the model group, the positive control group and 200 and 400 mg·L-1 DA groups showed prolonged paralysis time (P<0.05), and all the treatment groups showed decreased Aβ deposition in the head (P<0.01). DA within the concentration range of 50-500 mg·L-1 did not affect the viability of BV2 cells. In addition, DA enhanced the autophagy flux (P<0.05), up-regulated the mRNA levels of beclin1, Atg5, LAMP2, and CLN2 (P<0.05, P<0.01), promoted the nuclear translocation of TFEB (P<0.05), increased LAMP2 expression and autophagy flux (P<0.05, P<0.01), and enhanced the transcriptional activities of PPARα and TFEB (P<0.01). The positive control group and 200 and 400 mg·L-1 DA groups showed enhanced fluorescence intensity of PPARα in the BV2 nucleus (P<0.01). UPLC-MS detected nine known compounds of DA, from which 8 active components of DA were screened out. The docking results suggested that a variety of components in DA could bind to PPARα-LBD and form stable hydrogen bonds. ConclusionDA may reduce the pathological changes in AD by regulating the PPARα-TFEB pathway.
5.Distribution characteristics of IgA in Zhuang blood donors and its influence on blood transfusion safety in Nanning
Qiuhong MO ; Yang CHEN ; Mingshuang LAI ; Huihui MO ; Baoren HE ; Baojia HUANG ; Yanya TANG ; Linbin HUANG ; Limin CHEN
Chinese Journal of Blood Transfusion 2025;38(6):811-816
Objective: To analyze the distribution characteristics of immunoglobulin A (IgA) concentration in Nanning Zhuang blood donors by measuring the concentration of plasma IgA. Methods: Enzyme-linked immunosorbent assay (ELISA) was performed to measure the absorbance of 2 000 plasma samples from Zhuang blood donors. The IgA concentration in samples was calculated using the ELISA Calc regression/fitting technology program. Results: The standard curve demonstrated that ELISA detection of plasma IgA concentration exhibited good precision. The frequency of IgA deficiency was 0/2 000. No statistically significant difference in the distribution of IgA concentration was observed between males and females (P>0.05). The distribution of IgA concentration varied significantly across age groups: younger individuals (18-39 years old) had lower plasma IgA levels (mg/dL) compared to older individuals (40-56 years old): 5-89.99 mg/dL group, 8.80% (176/2 000) vs 17.20% (344/2 000); 90-450 mg/dL group,20.65% (413/2 000) vs 51.20% (1 024/2 000); >450 mg/dL group, 0.45%, (9/2 000) vs 1.70% (34/2 000), P<0.05. No significant difference in IgA concentration was found among different ABO blood types in Zhuang blood donors (P>0.05). Spearman correlation analysis revealed a positive correlation between age and IgA concentration (R
=0.114, P<0.05). Conclusion: No individuals with IgA deficiency were screened out among the Zhuang blood donors in Nanning area, and plasma IgA levels progressively increase with age.
6.Efficacy of implantable neuromuscular electrical stimulation system on stress urinary incontinence model in female rats
Bohong LONG ; Chen LI ; Han DENG ; Haoyu SUN ; Limin LIAO ; Xing LI
Journal of Modern Urology 2025;30(5):438-441
Objective: To investigate the efficacy of implantable neuromuscular electrical stimulation system on stress urinary incontinence (SUI) model in female rats. Methods: A total of 21 female infertile SD rats were randomly divided into the control,sham stimulation,and stimulation groups,with 7 rats in each group.All rats received vaginal dilation (VD) to simulate postpartum SUI.One week after VD,the control group was given normal feeding,stimulators were implanted in the pelvic floor muscles of the sham stimulation and stimulation groups.The sham stimulation group received normal feeding for 2 weeks,and the stimulation group received pelvic floor electrical stimulation (PFES) for 2 consecutive weeks.The leak point pressure (LPP) of each rat was measured with cystometry before VD (baseline value),1 week after VD,and 2 weeks after PFES. Results: In the control group and sham stimulation group,LPP increased after 2 weeks of treatment compared with that after 1 week of VD,but it still did not return to the baseline level (P<0.001).In the stimulation group,after 2 consecutive weeks of PFES,LPP increased significantly compared with that 1 week after VD,and returned to the baseline value (P>0.05).There was no significant difference in the LPP baseline values and levels after 1 week of VD among the 3 groups (P>0.05).The LPP in the stimulation group after 2 weeks of PFES was significantly higher than that in the sham stimulation group and stimulation group (P<0.001). Conclusion: The implantable neuromuscular electrical stimulation system is effective in short-term intervention of SUI in female rats,the further studies are needed to confirm the long-term efficacy and safety of the system,the optimal stimulation sites,optimal stimulation parameters,and potential mechanisms of action.
7.Research progress on periodontal pathogen inducing mitochondrial dysfunction promoting periodontitis
LI Limin ; PENG Xian ; ZHOU Xuedong
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(9):801-808
Mitochondria, ubiquitous energy-producing organelles in eukaryotic cells, can have their normal functions disrupted by bacterial infections, leading to mitochondrial dysfunction. This dysfunction is closely associated with inflammatory diseases. Periodontitis, a chronic inflammatory disorder of periodontal tissues caused by pathogenic microorganisms, has been increasingly linked to mitochondrial dysfunction in its pathogenesis and progression. Compared to healthy periodontal tissues, inflammatory lesions exhibit more pronounced mitochondrial dysfunction—a pathological process that is strongly correlated with periodontal pathogen infection. Studies reveal that these pathogens disrupt mitochondrial homeostasis in host cells (e.g., gingival epithelial cells and fibroblasts) through multiple mechanisms, including disrupting mitochondrial biogenesis, altering mitochondrial dynamics (promoting excessive fission), inhibiting mitophagy, impairing mitochondrial dysfunction-associated apoptosis, and inducing endogenous oxidative stress, which upregulates pro-inflammatory cytokines. Collectively, these processes drive the establishment and persistence of an inflammatory microenvironment. This review explores how periodontal pathogens affect mitochondrial function and their mechanistic contributions to periodontitis progression, with the goal of providing novel insights for developing mitochondria-targeted therapeutic strategies.
8.The influence of ASiR-V algorithm on radiation dose and image quality in children’s ultra-low-dose chest CT examination
Limin WANG ; Xiumin LYU ; Yanping WANG ; Lulu XU
Chinese Journal of Radiological Health 2025;34(4):540-545
Objective To explore the impact of multi-model adaptive statistical iterative reconstruction (ASiR-V) algorithm on radiation dose and image quality in children’s ultra-low-dose chest CT examination. Methods A total of 72 children who underwent chest CT scans at Qingdao Municipal Hospital with admissions between January 2024 and January 2025 were selected as subjects and divided into two groups using a random number table. In the control group (n = 36), the tube voltage was set at 100 kVp and the conventional filtered back projection algorithm was used. In the observation group (n = 36), the tube voltage was set at 80 kVp and images were reconstructed using 30% ASiR-V (observation group 1), 60% ASiR-V (observation group 2), and 90% ASiR-V (observation group 3), respectively. Radiation doses were recorded for each group, and both subjective and objective evaluations of image quality were conducted. Results Compared with the control group, the observation group demonstrated significantly lower volume CT dose index [(0.86 ± 0.09) mGy], dose length product [(25.90 ± 3.55) mGy·cm], and effective dose [(0.01 ± 0.001) mSv] (P < 0.05). There was no significant difference in subjective evaluation scores of image quality among the four groups (z = −2.206, P = 0.530). Additionally, Fisher’s exact test showed that the proportion of images scoring 4-5 points was higher in observation group 2 than in observation group 3 (P = 0.024). The noise value of the ascending aorta in the mediastinal window and the noise values of the right and left middle lung fields and the right and left upper lung fields in the lung window were lower in observation groups 2 and 3 than in the control group, and these values were lower in observation group 3 than in observation group 2 (P < 0.05). The signal-to-noise ratios of the ascending aorta and liver in observation groups 2 and 3 were higher than those in the control group, and the ratios were higher in observation group 3 than in observation group 2 (P < 0.05). Conclusion Reconstruction using the 60% ASiR-V algorithm for pediatric ultra-low-dose chest CT examination can ensure good image quality while reducing radiation dose and improving examination safety.
9.Shanxiangyuanye (Turpiniae Folium) for diabetic complications: chemical constituents and therapeutic potential
Ruiyao Xiong ; Shuang Chen ; Zihao Dai ; Limin Gong
Digital Chinese Medicine 2025;8(3):413-424
Objective:
To analyze the chemical constituents of Shanxiangyuanye (Turpiniae Folium) through liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, and to evaluate their anti-oxidant, hypoglycemic, and anti-glycation activities related to diabetic complications.
Methods:
The supernatant of Shanxiangyuanye (Turpiniae Folium) (TFS), obtained following water extraction and alcohol precipitation, was analyzed by LC-MS/MS. Antioxidant activity of TFS in vitro was evaluated using three experimental approaches: the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) radical cation decolorization assay, and the hydroxyl (·OH) radical scavenging assay. To comprehensively evaluate hypoglycemic potential, α-glucosidase inhibition was measured to analyze in vitro hypoglycemic activity. Subsequently, in vitro models were developed to examine anti-glycation activity through the bovine serum albumin (BSA)-fructose (Fru), BSA-methylglyoxal (MGO), BSA-glyoxal (GO), and D-arginine (Arg)-MGO systems, with particular attention to the inhibitory effects of TFS. Furthermore, the concentrations of fructosamine, protein carbonyls, sulfhydryl groups, and β-amyloid in the glycation solution were quantified using the BSA-Fru model following 7-d of incubation at 37 °C.
Results:
Using LC-MS/MS analysis in both positive and negative ion modes, we identified 750 chemical components in TFS, primarily including organic acids, amino acids, and their derivatives. In vitro activity studies demonstrated that TFS exhibited remarkable free radical scavenging capacity, with half-maximal inhibitory concentrations (IC50) of 0.47, 1.56, and 0.36 mg/mL against DPPH, ABTS+, and ·OH radicals, respectively. Regarding hypoglycemic activity, TFS dose-dependently inhibited α-glucosidase activity (IC50 = 0.21 mg/mL), displaying comparable efficacy to the clinical drug acarbose (IC50 = 0.23 mg/mL). Notably, TFS intervened in the glycation process: IC50 values were 0.22, 1.91 – 4.96, and 4.09 mg/mL in the BSA-Fru, BSA-MGO/GO, and Arg-MGO models, respectively, with the most prominent inhibitory effects observed in the BSA-Fru model. Furthermore, although TFS may not effectively preserve thiol groups in BSA or reduce thiol oxidation during glycation, it significantly reduces fructosamine levels (in a dose-dependent manner), decreases β-amyloid formation, and inhibits protein carbonylation (P < 0.000 1).
Conclusion
The findings demonstrate that TFS exhibits a complex chemical composition with potent antioxidant, hypoglycemic, and anti-glycation activities. These results provide compelling scientific evidence supporting TFS’s potential as a natural adjuvant for diabetes prevention and complication management, while laying a solid foundation for its applications in functional food development and adjunctive antidiabetic therapeutics.
10.EZH2 protein expression in predicting malignant transformation of oral leukoplakia: a prospective cohort study
MEI Nianrou ; LIU Limin ; YANG Jingwen ; XU Siming ; LI Chenxi ; GE Shuyun ; ZHOU Haiwen
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(10):862-872
Objective:
To investigate the predictive value of EZH2 expression for malignant transformation in oral leukoplakia (OLK) and to provide a reference for clinical practice.
Methods:
This study was approved by the institutional ethics committee, and informed consent was obtained from all participants. A total of 114 patients diagnosed with OLK by pathological examination and treated at our hospital between November 2020 and July 2022 were initially enrolled. After excluding those with incomplete data or follow-up, 105 participants were included in the final analysis, comprising 14 in the high EZH2 expression group and 91 in the low EZH2 expression group. Histopathological examination of oral mucosa and immunohistochemical detection of EZH2 protein expression were performed. The follow-up period was 30 months; participants were followed until malignant transformation occurred or until the end of follow-up, at which point they were withdrawn from the study. The exposure factor was the level of EZH2 protein expression, and the outcome was the malignant transformation rate of OLK. Differences in EZH2 expression levels and transformation outcomes were analyzed.
Results:
There were no statistically significant differences between the high and low EZH2 expression groups in terms of age, sex, history of systemic disease, lifestyle habits, psychological status, diet, and sleep conditions (P > 0.05). Lesions in the high EZH2 expression group were mainly located on the ventral tongue, while in the low EZH2 expression group, they were more commonly found on the dorsal tongue and buccal mucosa. The malignant transformation rate was 28.6% (4/14) in the high expression group and 8.8% (8/91) in the low expression group; these differences were not statistically significant (P=0.053). In univariate Cox regression analysis, the risk of malignant transformation in the high EZH2 expression group was 3.647 times that of the low EZH2 expression group (HR = 3.647, 95% CI: 1.097-12.120, P<0.05). Kaplan-Meier survival analysis showed that over the 30-month follow-up period, the cancer-free survival rate in the high EZH2 expression group was 19.8% lower than in the low expression group, and the difference was statistically significant (P<0.05). In multivariate Cox regression analysis, only moderate and severe epithelial dysplasia were identified as independent risk factors for malignant transformation. The risk of malignant transformation in the moderate and severe dysplasia groups was 10.695 and 13.623 times higher, respectively, than in the mild dysplasia group (HR = 10.695, 95% CI: 2.270-50.396, P<0.05; HR=13.623, 95% CI: 1.918-96.774, P<0.05). EZH2 high expression was not an independent risk factor in the multivariate model (HR= 2.528, 95% CI: 0.752-8.500, P = 0.134).
Conclusion
High EZH2 protein expression is a risk factor for the malignant transformation of OLK but does not have independent predictive value.


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